Are there any types of cylindrical cell culture inserts that are designed to confine cell seeding area? Say, if I'm using a 12 well plate (3.5 cm^2/well), but only want to seed cells in the middle 1.9 cm^2 area. If so, are there any reusable insert options or only disposable ones? Thanks in advance.
You can try adding a drop of medium in the centre of the well (Do not pre-wet the well before adding your drop!). Try different volumes so that you can obtain your desired sureface area.
After knowing the volume, prepare your cell suspension at a proper cencentration and add drops onto your plate. Avoid tilting or disturbing the plate too much so that the droplet can maintain the hemisphere shape. Wait until the cells adhere to the surface and you can replenish enough growth medium to the wells.
Hi Matthew Huang
You could try coating a glass coverslip with an adhesive layer, seed your cells onto that and then pick up the coverslip using sterile forceps and transfer to a fresh well.
there are multiple companies that make cloning cylinders that work for this purpose. Example : https://www.fishersci.com/shop/products/cloning-cylinder-10x10mm-pk125/501532241. THey come in many sizes. If you have access, a glassblowing shop can custom-make some of other sizes from pyrex tubing.
Hi Matthew,
From your description it is not clear what your cells grown in the centre of the wells will be used for. As such, it is difficult to suggest which technique/approach would work for you the best. All suggestions provided above are excellent ones, however, they will not prevent cells from growing/migrating outwards from the seeding area. If your experiment requires maintaining the cells in a very confined area on a 35mm dish, I would recommend to check "micro-Dish (u-Dish)" and/or "Culture-Insert in u-Dish" products from "ibidi" that offer a range of different options for growing the cells on a different confined area(s) in the middle/center of the dish.
Best of luck
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